Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis
Summary: Patient-derived organoids (PDOs) are ideal ex vivo model systems to study cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring drug efficacy in three-dimensional (3D) high-grade serous ovarian cancer PDO cultures through quantification of cytotoxicity...
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006500 |
| _version_ | 1797615910121373696 |
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| author | Aikaterini Skorda Anna Røssberg Lauridsen Kaisa Huhtinen Alexandra Lahtinen Wojciech Senkowski Jaana Oikkonen Johanna Hynninen Sampsa Hautaniemi Tuula Kallunki |
| author_facet | Aikaterini Skorda Anna Røssberg Lauridsen Kaisa Huhtinen Alexandra Lahtinen Wojciech Senkowski Jaana Oikkonen Johanna Hynninen Sampsa Hautaniemi Tuula Kallunki |
| author_sort | Aikaterini Skorda |
| collection | DOAJ |
| description | Summary: Patient-derived organoids (PDOs) are ideal ex vivo model systems to study cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring drug efficacy in three-dimensional (3D) high-grade serous ovarian cancer PDO cultures through quantification of cytotoxicity using propidium iodide incorporation in dead cells. We also provide detailed steps to analyze proliferation of PDOs using the Ki67 biomarker. We describe steps for sample processing, immunofluorescent staining, high-throughput confocal imaging, and image-based quantification for 3D cultures.For complete details on the use and execution of this protocol, please refer to Lahtinen et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-11T07:33:41Z |
| format | Article |
| id | doaj.art-330b361b02744b8a9f7f5805c6c8fd54 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-11T07:33:41Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-330b361b02744b8a9f7f5805c6c8fd542023-11-17T05:28:10ZengElsevierSTAR Protocols2666-16672023-12-0144102683Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysisAikaterini Skorda0Anna Røssberg Lauridsen1Kaisa Huhtinen2Alexandra Lahtinen3Wojciech Senkowski4Jaana Oikkonen5Johanna Hynninen6Sampsa Hautaniemi7Tuula Kallunki8Danish Cancer Institute, Danish Cancer Society, 2100 Copenhagen, DenmarkDanish Cancer Institute, Danish Cancer Society, 2100 Copenhagen, DenmarkResearch Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland; Institute of Biomedicine and FICAN West Cancer Centre, University of Turku and Turku University Hospital, 20014 Turku, FinlandResearch Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, FinlandBiotech Research and Innovation Centre, University of Copenhagen, 2200 Copenhagen, DenmarkResearch Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, FinlandDepartment of Obstetrics and Gynaecology, University of Turku and Turku University Hospital, 200521 Turku, FinlandResearch Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, FinlandDanish Cancer Institute, Danish Cancer Society, 2100 Copenhagen, Denmark; Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark; Corresponding authorSummary: Patient-derived organoids (PDOs) are ideal ex vivo model systems to study cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring drug efficacy in three-dimensional (3D) high-grade serous ovarian cancer PDO cultures through quantification of cytotoxicity using propidium iodide incorporation in dead cells. We also provide detailed steps to analyze proliferation of PDOs using the Ki67 biomarker. We describe steps for sample processing, immunofluorescent staining, high-throughput confocal imaging, and image-based quantification for 3D cultures.For complete details on the use and execution of this protocol, please refer to Lahtinen et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723006500Cell BiologyCancerHigh-Throughput ScreeningOrganoids |
| spellingShingle | Aikaterini Skorda Anna Røssberg Lauridsen Kaisa Huhtinen Alexandra Lahtinen Wojciech Senkowski Jaana Oikkonen Johanna Hynninen Sampsa Hautaniemi Tuula Kallunki Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis STAR Protocols Cell Biology Cancer High-Throughput Screening Organoids |
| title | Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis |
| title_full | Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis |
| title_fullStr | Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis |
| title_full_unstemmed | Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis |
| title_short | Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis |
| title_sort | quantification of cell death and proliferation of patient derived ovarian cancer organoids through 3d imaging and image analysis |
| topic | Cell Biology Cancer High-Throughput Screening Organoids |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006500 |
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